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Enhanced Production of Recombinant Alcohol Dehydrogenase Using the Genetically Engineered Escherichia Coli Strain That Heterologously Expresses Carrot Heat Shock Protein 70

Enhanced Production of Recombinant Alcohol Dehydrogenase Using the Genetically Engineered Escherichia Coli Strain That Heterologously Expresses Carrot Heat Shock Protein 70

Minhye Kim, Bomin Jang, Yeh-Jin Ahn

Curr Microbiol. 2019 Nov;76(11):1338-1344.

PMID: 31254010

Abstract:

Escherichia coli (E. coli) has been widely used as a host organism for producing recombinant proteins such as biocatalysts, antibody fragments, and therapeutic hormones. To enhance recombinant protein production, many E. coli strains have been genetically engineered on practical purposes. In this study, we developed the engineered E. coli strain expressing Heat shock protein 70, DcHsp70, from carrot (Daucus carota L.). The DNA construct for DcHsp70 expression, Lipoprotein promoter-DcHsp70 gene-Flippase recognition target cassette, which is flanked by the insertion site yddE pseudogene sequences, was generated by overlap PCR and inserted into the E. coli genome by lambda Red-mediated homologous recombination. To examine if the engineered E. coli cells can effectively produce recombinant proteins, the alcohol dehydrogenase (ADH) gene from a thermophile, Geobacillus stearothermophilus, was cloned into a pET11a expression vector and expressed by isopropyl β-D-1-thiogalactopyranoside treatment. Compared to wild type, the genetically engineered E. coli expressing DcHsp70 exhibited up to approximately 11-fold higher production of his-tagged ADH, mostly in soluble forms. The his-ADH protein that was purified from the engineered cells exhibited the enzyme activity. The genetically engineered E. coli developed in this study can be useful for the efficient production of recombinant proteins, such as recombinant ADH.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP9028120	Alcohol Dehydrogenase, recombinant from E. coli		9028-12-0	Price

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